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重组超氧化物歧化酶C是一种通过酶联免疫吸附测定法诊断伪结核棒状杆菌的潜在抗原。

rSodC is a potential antigen to diagnose Corynebacterium pseudotuberculosis by enzyme-linked immunoassay.

作者信息

Farias Antonio Pedro Fróes de, Rocha Filho José Tadeu Raynal, Marchioro Silvana Beutinger, Moreira Luan Santana, Marques Andressa Souza, Sá Maria da Conceição Aquino de, Oliveira Antonio Anderson Dos Santos, Alcântara Maria Emília, Mariutti Ricardo Barros, Arni Raghuvir Krishnaswamy, Trindade Soraya Castro, Meyer Roberto

机构信息

Immunology and Molecular Biology Laboratory (LABIMUNO), Federal University of Bahia (UFBA), Salvador, BA, Brazil.

College of Science and Entrepreneurship (FACEMP), Santo Antônio de Jesus, BA, Brazil.

出版信息

AMB Express. 2020 Oct 19;10(1):186. doi: 10.1186/s13568-020-01125-0.

Abstract

Caseous lymphadenitis (CL) is a chronic infectious disease that affects sheep and goats. Many serological tests have been developed to detect the disease; one of the most widely used is the enzyme-linked immunosorbent assay (ELISA), due to its advantages, which include acceptable cost-effectiveness, applicability, sensitivity and specificity. ELISA formulations using recombinant proteins can exhibit significant sensitivity and specificity when using a single purified antigen. DTxR, Trx, TrxR, LexA, SodC, SpaC, NanH, and PknG recombinant proteins can be considered target proteins for ELISA development due to its extracellular or on the cell surface location, which allows a better recognition by the immune system. Therefore, the objectives of this study were to evaluate the antigenic reactivity of Corynebacterium pseudotuberculosis recombinant proteins in goat and sheep serum. Of eight proteins evaluated, rSodC was selected for validation assays with small ruminant serum samples from the semiarid region of the state of Bahia, Brazil. Validation assays with goat serum samples showed that ELISA-rSodC presented sensitivity and specificity of 96% and 94%, respectively. Validation assays with sheep serum showed that ELISA-rSodC exhibited sensitivity and specificity of 95% and 98%, respectively. Analysis of 756 field serum samples showed that rSodC identified 95 positive samples (23%) in goats and 75 positive samples (21%) in sheep. The ELISA with recombinant SodC protein developed in this study discriminated positive and negative serum samples with high levels of sensitivity and specificity. This formulation is promising for epidemiological surveys and CL control programs.Trial registration AEC No 4958051018. 12/18/2018, retrospectively registered.

摘要

干酪性淋巴结炎(CL)是一种影响绵羊和山羊的慢性传染病。已经开发了许多血清学检测方法来检测这种疾病;其中使用最广泛的一种是酶联免疫吸附测定(ELISA),因为它具有成本效益可接受、适用性强、灵敏度和特异性高等优点。使用重组蛋白的ELISA制剂在使用单一纯化抗原时可表现出显著的灵敏度和特异性。由于DTxR、Trx、TrxR、LexA、SodC、SpaC、NanH和PknG重组蛋白位于细胞外或细胞表面,能被免疫系统更好地识别,因此可被视为ELISA开发的靶蛋白。因此,本研究的目的是评估伪结核棒状杆菌重组蛋白在山羊和绵羊血清中的抗原反应性。在评估的8种蛋白质中,选择rSodC用于对来自巴西巴伊亚州半干旱地区的小反刍动物血清样本进行验证试验。用山羊血清样本进行的验证试验表明,ELISA-rSodC的灵敏度和特异性分别为96%和94%。用绵羊血清进行的验证试验表明,ELISA-rSodC的灵敏度和特异性分别为95%和98%。对756份现场血清样本的分析表明,rSodC在山羊中鉴定出95份阳性样本(23%),在绵羊中鉴定出75份阳性样本(21%)。本研究中开发的重组SodC蛋白ELISA能够以高灵敏度和特异性区分阳性和阴性血清样本。这种制剂在流行病学调查和CL控制计划方面很有前景。试验注册AEC No 4958051018。2018年12月18日,追溯注册。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0930/7573032/79fb3468c27a/13568_2020_1125_Fig1_HTML.jpg

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